CN107711615A - A kind of method that sterile fish is obtained using immersion fish-egg - Google Patents
A kind of method that sterile fish is obtained using immersion fish-egg Download PDFInfo
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- CN107711615A CN107711615A CN201711049382.7A CN201711049382A CN107711615A CN 107711615 A CN107711615 A CN 107711615A CN 201711049382 A CN201711049382 A CN 201711049382A CN 107711615 A CN107711615 A CN 107711615A
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- sterile
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- 238000000034 method Methods 0.000 title claims abstract description 48
- 241000251468 Actinopterygii Species 0.000 title claims abstract description 27
- 238000007654 immersion Methods 0.000 title claims abstract description 19
- 230000012447 hatching Effects 0.000 claims abstract description 36
- 230000009027 insemination Effects 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 23
- 201000010099 disease Diseases 0.000 claims abstract description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 12
- 210000000582 semen Anatomy 0.000 claims abstract description 10
- 238000009395 breeding Methods 0.000 claims abstract description 7
- 230000001488 breeding effect Effects 0.000 claims abstract description 7
- 230000001629 suppression Effects 0.000 claims abstract description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 56
- WVLHHLRVNDMIAR-IBGZPJMESA-N AMD 070 Chemical compound C1CCC2=CC=CN=C2[C@H]1N(CCCCN)CC1=NC2=CC=CC=C2N1 WVLHHLRVNDMIAR-IBGZPJMESA-N 0.000 claims description 24
- 229940121384 cxc chemokine receptor type 4 (cxcr4) antagonist Drugs 0.000 claims description 23
- 235000013601 eggs Nutrition 0.000 claims description 23
- 230000002710 gonadal effect Effects 0.000 claims description 10
- 102000002322 Egg Proteins Human genes 0.000 claims description 9
- 108010000912 Egg Proteins Proteins 0.000 claims description 9
- 210000004681 ovum Anatomy 0.000 claims description 9
- 208000027418 Wounds and injury Diseases 0.000 claims description 6
- 210000001015 abdomen Anatomy 0.000 claims description 6
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 6
- 208000014674 injury Diseases 0.000 claims description 6
- 230000035800 maturation Effects 0.000 claims description 6
- 239000012528 membrane Substances 0.000 claims description 6
- 229910052760 oxygen Inorganic materials 0.000 claims description 6
- 239000001301 oxygen Substances 0.000 claims description 6
- 230000006835 compression Effects 0.000 claims description 5
- 238000007906 compression Methods 0.000 claims description 5
- 239000007788 liquid Substances 0.000 claims description 3
- 230000033228 biological regulation Effects 0.000 claims 1
- 230000008901 benefit Effects 0.000 abstract description 10
- 230000018109 developmental process Effects 0.000 abstract description 7
- 230000004083 survival effect Effects 0.000 abstract description 5
- 102100031650 C-X-C chemokine receptor type 4 Human genes 0.000 abstract description 4
- 101000922348 Homo sapiens C-X-C chemokine receptor type 4 Proteins 0.000 abstract description 4
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 7
- 102100021669 Stromal cell-derived factor 1 Human genes 0.000 description 5
- 101710088580 Stromal cell-derived factor 1 Proteins 0.000 description 5
- 230000004720 fertilization Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 108010061299 CXCR4 Receptors Proteins 0.000 description 4
- 102000012000 CXCR4 Receptors Human genes 0.000 description 4
- UELITFHSCLAHKR-UHFFFAOYSA-N acibenzolar-S-methyl Chemical compound CSC(=O)C1=CC=CC2=C1SN=N2 UELITFHSCLAHKR-UHFFFAOYSA-N 0.000 description 4
- RGXWDWUGBIJHDO-UHFFFAOYSA-N ethyl decanoate Chemical compound CCCCCCCCCC(=O)OCC RGXWDWUGBIJHDO-UHFFFAOYSA-N 0.000 description 4
- 208000000509 infertility Diseases 0.000 description 4
- 230000036512 infertility Effects 0.000 description 4
- 231100000535 infertility Toxicity 0.000 description 4
- 230000019491 signal transduction Effects 0.000 description 4
- 231100000331 toxic Toxicity 0.000 description 3
- 230000002588 toxic effect Effects 0.000 description 3
- 150000003935 benzaldehydes Chemical class 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 102000009410 Chemokine receptor Human genes 0.000 description 1
- 108050000299 Chemokine receptor Proteins 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 239000002576 chemokine receptor CXCR4 antagonist Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 150000003462 sulfoxides Chemical class 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
- A01K61/17—Hatching, e.g. incubators
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses a kind of method for obtaining sterile fish using immersion fish-egg, including parent culture, the collection of fish-egg and seminal fluid, artificial insemination, embryonated egg suppression processing, hatch and cultivate.Have the beneficial effect that：A kind of method that sterile fish is obtained using immersion fish-egg provided by the invention can upset the α CXCR4 signal paths of SDF 1, suppress the development of embryonic period, embryonic phase reproduction Yuan bases, have huge social benefit, economic benefit and ecological benefits；Suitable breeding density can improve hatching survival rate with appropriate hatching barrel water velocity, shorten brooding time, and the fry quality better hatched, deformity is few, and disease immune ability is strong.
Description
Technical field
The present invention relates to technical field of fishery, more particularly, to a kind of method for obtaining sterile fish using immersion fish-egg.
Background technology
Sterile fish has higher scientific research and application value in fishery, and first, sterile fish shows to grow fast, dressing percentage
High advantage；Secondly, sterile fish can reduce the ecological risk for escaping into nature by fish and bringing；Furthermore sterile fish is in section
There is special application, for example in Technique in Fishes replace-conceive research, sterile fish produces the sperm of other fingerlings as acceptor in grinding
Or ovum.Therefore, the method for exploring the sterile fish of generation is significant in fishery field.
The content of the invention
It is an object of the invention to provide a kind of method for obtaining sterile fish using immersion fish-egg, this method is simple, operation
It is convenient, SDF-1 α-CXCR4 signal paths can be upset, suppress the development of embryonic period, embryonic phase reproduction Yuan bases, and toxic side effect is faint, tool
There are huge social benefit, economic benefit and ecological benefits.
The technical scheme taken to achieve the above object of the present invention is：A kind of side that sterile fish is obtained using immersion fish-egg
Method, comprise the following steps：
Parent culture：Physical strong, disease-free and injury-free raun and milter is selected to carry out reinforced cultivating respectively as parent, cultivate
Water temperature is 19-22 DEG C, and salinity be 30-32 ‰, dissolved oxygen 5.3-6mg/L, should under the conditions of, cultivation water body, bion and
Microecosystem is in dynamic equilibrium all the time, establishes benign ecological environment space for parent, reached health, safety,
It is efficient to cultivate purpose；
The collection of fish-egg and seminal fluid：Parent culture gathers fish-egg and seminal fluid to gonadal maturation, using manual compression belly method,
Good condition is provided for artificial insemination by the step；
Artificial insemination：Dry insemination method carries out artificial insemination, is inseminated using dry method, step process is few, and artificial insemination time is short, and energy
Enough reach and meet expected rate of fertilization；
Embryonated egg suppression is handled：AMD070 CXCR4 antagonists are mixed with dimethyl sulfoxide (DMSO), into gained mixed liquor add by
Smart ovum soaks 24-48h, AMD070 CXCR4 antagonists wherein in the mixed liquor of AMD070 CXCR4 antagonists and dimethyl sulfoxide (DMSO)
Mass fraction be 0.1-0.3%, method is simple, easy to operate, should under the conditions of, AMD070 CXCR4 antagonists can upset SDF-
1 α-CXCR4 signal paths, suppress CXCR4 receptor signaling pathways, and then the development of embryonic period, embryonic phase Gonadal anlage can be suppressed, so as to
The effect of infertility is reached, and toxic side effect is faint；
Hatching and cultivation：Embryonated egg, which is placed in hatching barrel, hatches, and hatching density is 190-200g embryonated eggs/m3, hatching barrel water
Flow velocity is 3-4cm/s, and to prelarva i.e. by membrane, floating ovum is moved into nursery pond culture, should under the conditions of, suitable breeding density with
Appropriate hatching barrel water velocity can improve hatching survival rate, shorten brooding time, and the fry quality better hatched, deformity
Few, disease immune ability is strong.
Compared with prior art, beneficial effects of the present invention are：A kind of use provided by the invention soaks fish-egg acquisition not
SDF-1 α-CXCR4 signal paths can be upset by educating the method for fish, suppress the development of embryonic period, embryonic phase reproduction Yuan bases, have huge society
Can benefit, economic benefit and ecological benefits；Suitable breeding density and appropriate hatching barrel water velocity can improve hatching into
Motility rate, shorten brooding time, and the fry quality better hatched, deformity is few, and disease immune ability is strong.
Embodiment
It is described in further detail with reference to embodiments：
Embodiment 1：A kind of method for obtaining sterile fish using immersion fish-egg, comprises the following steps：
1) parent culture：Physical strong, disease-free and injury-free raun and milter is selected to carry out reinforced cultivating respectively as parent, train
Water temperature is educated for 19 DEG C, and salinity is 30 ‰, dissolved oxygen 5.3mg/L, under the conditions of being somebody's turn to do, cultivates water body, bion and Tiny ecosystem
System is in dynamic equilibrium all the time, is established benign ecological environment space for parent, has been reached healthy, safe and efficient
Cultivate purpose；
2) collection of fish-egg and seminal fluid：Parent culture gathers fish-egg and essence to gonadal maturation using manual compression belly method
Liquid, good condition is provided for artificial insemination by the step；
3) artificial insemination：Dry insemination method carries out artificial insemination, is inseminated using dry method, and step process is few, and artificial insemination time is short, and
It can reach and meet expected rate of fertilization；
4) embryonated egg suppression is handled：AMD070 CXCR4 antagonists are mixed with dimethyl sulfoxide (DMSO), added into gained mixed liquor
Embryonated egg soaks 24h, AMD070 CXCR4 antagonists wherein in the mixed liquor of AMD070 CXCR4 antagonists and dimethyl sulfoxide (DMSO)
Mass fraction be 0.1%, method is simple, easy to operate, should under the conditions of, AMD070 CXCR4 antagonists can upset SDF-1 α-
CXCR4 signal paths, suppress CXCR4 receptor signaling pathways, and then the development of embryonic period, embryonic phase Gonadal anlage can be suppressed, so as to reach
The effect of infertility；
5) hatch and cultivate：Embryonated egg, which is placed in hatching barrel, hatches, and hatching density is 190g embryonated eggs/m3, hatching barrel water stream
Speed is 3cm/s, and to prelarva i.e. by membrane, floating ovum is moved into nursery pond culture, should under the conditions of, suitable breeding density with it is appropriate
Hatching barrel water velocity can improve hatching survival rate, shorten brooding time, and the fry quality better hatched, deformity is few,
Disease immune ability is strong.
Embodiment 2：A kind of method for obtaining sterile fish using immersion fish-egg, comprises the following steps：It is parent culture first：
Physical strong, disease-free and injury-free raun and milter is selected to carry out reinforced cultivating respectively, it is 22 DEG C to cultivate water temperature, salt as parent
Spend for 32 ‰, dissolved oxygen 6mg/L；Followed by the collection of fish-egg and seminal fluid：Parent culture is to gonadal maturation, using artificial
Extrude belly method collection fish-egg and seminal fluid：Dry insemination method carries out artificial insemination, is inseminated using dry method, step process is few, manually awards
The smart time is short, and can reach and meet expected rate of fertilization；Handled followed by embryonated egg suppression：By AMD070 CXCR4 antagonists
Mixed with dimethyl sulfoxide (DMSO), embryonated egg immersion 48h, wherein AMD070 CXCR4 antagonists and diformazan are added into gained mixed liquor
The mass fraction of AMD070 CXCR4 antagonists is 0.3% in the mixed liquor of base sulfoxide；It is finally hatching and cultivation：Embryonated egg is put
Hatch in hatching barrel, hatching density is 200g embryonated eggs/m3, hatching barrel water flow velocity be 4cm/s, to prelarva i.e. by membrane,
Floating ovum is moved into nursery pond culture.
Embodiment 3：A kind of method for obtaining sterile fish using immersion fish-egg, comprises the following steps：
1) parent culture：Physical strong, disease-free and injury-free raun and milter is selected to carry out reinforced cultivating respectively as parent, train
Water temperature is educated for 21 DEG C, and salinity is 31 ‰, dissolved oxygen 5.5mg/L, under the conditions of being somebody's turn to do, cultivates water body, bion and Tiny ecosystem
System is in dynamic equilibrium all the time, is established benign ecological environment space for parent, has been reached healthy, safe and efficient
Cultivate purpose；
2) collection of fish-egg and seminal fluid：Parent culture gathers fish-egg and essence to gonadal maturation using manual compression belly method
Liquid, good condition is provided for artificial insemination by the step；
3) artificial insemination：Dry insemination method carries out artificial insemination, is inseminated using dry method, and step process is few, and artificial insemination time is short, and
It can reach and meet expected rate of fertilization；
4) embryonated egg suppression is handled：AMD070 CXCR4 antagonists are mixed with dimethyl sulfoxide (DMSO), added into gained mixed liquor
Embryonated egg soaks 40h, AMD070 CXCR4 antagonists wherein in the mixed liquor of AMD070 CXCR4 antagonists and dimethyl sulfoxide (DMSO)
Mass fraction be 0.2%, method is simple, easy to operate, should under the conditions of, AMD070 CXCR4 antagonists can upset SDF-1 α-
CXCR4 signal paths, suppress CXCR4 receptor signaling pathways, and then the development of embryonic period, embryonic phase Gonadal anlage can be suppressed, so as to reach
The effect of infertility；
5) hatch and cultivate：Embryonated egg, which is placed in hatching barrel, hatches, and hatching density is 195g embryonated eggs/m3, hatching barrel water stream
Speed is 3cm/s, and to prelarva i.e. by membrane, floating ovum is moved into nursery pond culture, should under the conditions of, suitable breeding density with it is appropriate
Hatching barrel water velocity can improve hatching survival rate, shorten brooding time, and the fry quality better hatched, deformity is few,
Disease immune ability is strong.
Embodiment 4：A kind of method for obtaining sterile fish using immersion fish-egg, comprises the following steps：It is parent culture first：
Physical strong, disease-free and injury-free raun and milter is selected to carry out reinforced cultivating respectively, it is 21 DEG C to cultivate water temperature, salt as parent
Spend for 31 ‰, dissolved oxygen 5.6mg/L, should under the conditions of, cultivate water body, bion and microecosystem all the time in dynamic
In balance, benign ecological environment space is established for parent, has reached healthy, safe and efficient cultivation purpose；Followed by fish
The collection of ovum and seminal fluid：Parent culture gathers fish-egg and seminal fluid, by this to gonadal maturation using manual compression belly method
Step provides good condition for artificial insemination；Followed by artificial insemination：Dry insemination method carries out artificial insemination, is awarded using dry method
Essence, step process is few, and artificial insemination time is short, and can reach and meet expected rate of fertilization；Handled followed by embryonated egg suppression：
AMD070 CXCR4 antagonists are mixed with dimethyl sulfoxide (DMSO), 10min is stirred, adds and account for dimethyl sulfoxide (DMSO) quality 0.1 ‰
3- trifluoromethylated benzaldehydes and the ethyl caprate for accounting for dimethyl sulfoxide (DMSO) quality 0.3 ‰, 15min is stirred, added into gained mixed liquor
Enter embryonated egg immersion 40h, AMD070 CXCR4 antagonisms wherein in the mixed liquor of AMD070 CXCR4 antagonists and dimethyl sulfoxide (DMSO)
The mass fraction of agent is 0.25%, and method is simple, easy to operate, and under the conditions of being somebody's turn to do, AMD070 CXCR4 antagonists can upset SDF-1
α-CXCR4 signal paths, suppress CXCR4 receptor signaling pathways, and then the development of embryonic period, embryonic phase Gonadal anlage can be suppressed, so as to reach
The effect of infertility is arrived, 3- trifluoromethylated benzaldehydes, ethyl caprate and the dimethyl sulfoxide (DMSO) added has synergy, increases
Other chemokine receptors are nearly free from inhibitory action by the strong selectivity of AMD070 CXCR4 antagonist inhibitory action,
It reduce further the toxic side effect of AMD070 CXCR4 antagonists；Followed by hatching and cultivate：Embryonated egg is placed in hatching barrel
Hatching, hatching density is 195g embryonated eggs/m3, hatching barrel water flow velocity be 3cm/s, to prelarva i.e. by membrane, floating ovum is moved
To nursery pond culture, under the conditions of being somebody's turn to do, suitable breeding density can improve hatching survival rate with appropriate hatching barrel water velocity,
Shorten brooding time, and the fry quality better hatched, deformity is few, and disease immune ability is strong.
Embodiment of above is merely to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this area
Member, without departing from the spirit and scope of the present invention, it can also make a variety of changes and modification.Therefore, it is all equivalent
Technical scheme fall within scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (6)
- A kind of 1. method that sterile fish is obtained using immersion fish-egg, it is characterised in that：It the described method comprises the following steps：1) parent culture：Physical strong, disease-free and injury-free raun and milter is selected to carry out reinforced cultivating respectively as parent；2) collection of fish-egg and seminal fluid：Parent culture gathers fish-egg and essence to gonadal maturation using manual compression belly method Liquid；3) artificial insemination：Dry insemination method carries out artificial insemination；4) embryonated egg suppression is handled：AMD070 CXCR4 antagonists are mixed with dimethyl sulfoxide (DMSO), added into gained mixed liquor Embryonated egg is soaked；5) hatch and cultivate：Embryonated egg, which is placed in hatching barrel, hatches, and the flow velocity of regulation hatching density and hatching barrel water, is to prelarva By membrane, floating ovum is moved into nursery pond culture.
- A kind of 2. method that sterile fish is obtained using immersion fish-egg according to claim 1, it is characterised in that：The step 1) breeding condition is in：It is 19-22 DEG C to cultivate water temperature, and salinity is 30-32 ‰, dissolved oxygen 5.3-6mg/L.
- A kind of 3. method that sterile fish is obtained using immersion fish-egg according to claim 1, it is characterised in that：The step The mass fraction of AMD070 CXCR4 antagonists is in the mixed liquor of rapid 4) AMD070 CXCR4 antagonists and dimethyl sulfoxide (DMSO) 0.1-0.3%.
- A kind of 4. method that sterile fish is obtained using immersion fish-egg according to claim 1, it is characterised in that：The step 4) soak time is 24-48h in.
- A kind of 5. method that sterile fish is obtained using immersion fish-egg according to claim 1, it is characterised in that：The step 5) hatching density is 190-200g embryonated eggs/m in3。
- A kind of 6. method that sterile fish is obtained using immersion fish-egg according to claim 1, it is characterised in that：The step 5) flow velocity of hatching barrel water is 3-4cm/s in.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711049382.7A CN107711615B (en) | 2017-10-31 | 2017-10-31 | Method for obtaining sterile fish by soaking fish eggs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711049382.7A CN107711615B (en) | 2017-10-31 | 2017-10-31 | Method for obtaining sterile fish by soaking fish eggs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107711615A true CN107711615A (en) | 2018-02-23 |
| CN107711615B CN107711615B (en) | 2020-08-11 |
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| CN201711049382.7A Expired - Fee Related CN107711615B (en) | 2017-10-31 | 2017-10-31 | Method for obtaining sterile fish by soaking fish eggs |
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|---|---|---|---|---|
| CN1733908A (en) * | 2005-07-05 | 2006-02-15 | 大连水产学院 | Abalone triploid pharmaceutical induction method |
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| CN106106279A (en) * | 2016-06-30 | 2016-11-16 | 湖南文理学院 | A kind of raising bream, Carnis Megalobramae triploid incubation rate method |
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-
2017
- 2017-10-31 CN CN201711049382.7A patent/CN107711615B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1733908A (en) * | 2005-07-05 | 2006-02-15 | 大连水产学院 | Abalone triploid pharmaceutical induction method |
| CN1957686A (en) * | 2006-11-20 | 2007-05-09 | 湖南文理学院 | Method of breeding pearls by using 6-dimethylin purine to induce triploid jibheader clams |
| CN102695798A (en) * | 2009-11-23 | 2012-09-26 | 水慷科技公司 | Maternally induced sterility in animals |
| CN105431521A (en) * | 2013-02-28 | 2016-03-23 | 哈佛学院校长同事会 | Methods and compositions for mobilizing stem cells |
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Granted publication date: 20200811 |